牙本质非胶原蛋白对人根尖牙乳头干细胞增殖及分化能力的影响

The effects of dentin non-collagenous proteins on the proliferation and mineralization of human stem cells from the apical papilla

目的:探讨牙本质非胶原蛋白(DNCPs)对人根尖牙乳头干细胞(h SCAPs)增殖及分化能力的影响。方法:h SCAPs经DNCPs诱导后,用MTT法检测细胞增殖能力;碱性磷酸酶(ALP)试剂盒测定ALP;RT-PCR检测ALP、骨钙素(OCN)及I型胶原(Col I)的m RNA表达变化;茜素红矿化结节染色及定量检测DNCPs对h SCAPs分化能力的影响。结果:DNCPs诱导1、3、5、7 d时,h SCAPs的增殖活性与对照组相比无显著差异(P>0.05);诱导7、14 d时,h SCAPs的ALP、Col I和OCN m RNA表达水平均显著上调(P<0.05);诱导3、5、7 d时,h SCAPs的ALP活性明显上调(P<0.05);诱导3周后,茜素红染色显示DNCPs诱导组的钙化结节形成能力明显高于对照组(P<0.05)。结论:DNCPs可明显促进h SCAPs矿化能力,对其增殖活性的影响则不明显。

AIM： To investigate the effects of dentin non-collagenous proteins（ DNCPs） on the proliferation and differentiation of human stem cells from the apical papilla（ h SCAPs） in vitro. METHODS： h SCAPs were treated with 10 μg / m L of DNCPs,and the proliferation of the h SCAPs was detected by MTT assay. To investigate the differentiation of h SCAPs,alkaline phosphotase（ ALP） activity was determined by enzyme kinetics methods,the m RNA expression of ALP,OCN and COL I was detected by real-time PCR,and the mineralization was detected by Alizarin red staining assay. RESULTS： The proliferation activity was not statistically different differect between the DNCPs- treated,and the control cells（ P〉0. 05）. After 7 and 14 days culture in DNCPs containing medium,the m RNA expressions of ALP,OCN and COL I in h SCAPs were significantly upregulated（ P〈0. 05）. ALP activity of d NCPs- treated cells was significantly promoted（ P〈0. 05）. After 3 weeks,the calcified nodules in DNCPs-treated cells were bigger and more that those in the controls（ P〈0. 05）. CONCLUSION： DNCPs can promote the mineralization ability of h SCAPs,while have no significant effect on the proliferation ability.